Development of optical hypoxic environment for manufacturing a tissue-engineered oral mucosa by utilizing a hypoxia-sensing luminescent probe

• Project/Area Number: 26293420
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Dental engineering/Regenerative dentistry
• Research Institution: Niigata University
• Principal Investigator: IZUMI KENJI 新潟大学, 医歯学系, 教授 (80242436)
• Co-Investigator(Kenkyū-buntansha): 飛田 成史 群馬大学, 大学院理工学府, 教授 (30164007) ; 前田 健康 新潟大学, 医歯学系, 教授 (40183941)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥16,120,000 (Direct Cost: ¥12,400,000、Indirect Cost: ¥3,720,000); Fiscal Year 2016: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2014: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
• Keywords: 低酸素 / 再生医療 / 口腔粘膜 / 角化細胞 / 幹細胞ニッチ / 口腔粘膜角化細胞 / 発光プローブ / 口腔粘膜上皮細胞 / 上皮細胞 / プローブ

Outline of Final Research Achievements

It is necessary to calibrate cellular oxygen levels to quantify oxygen levels of an epithelial layer within a tissue-engineered oral mucosa for clinical application. In this study, we used a new iridium complex BTPDM1 as an oxygen sensing probe, at a concentration of 500nM. When culturing oral keratinocytes in a monolayer, in a variety of culture conditions such as cell plating density and calcium concentration, we measured intracellular oxygen levels using confocal phosphorescence lifetime imaging microscope. Our results showed cellular oxygen levels appeared to be dependent on the size of cells, and the development of cornified layer did not cause severe hypoxic condition in the underlying cell layer. This suggests the intraepithelial oxygen levels of the EVPOME grafts may depend on the cellular differentiation and the quality of cornified layer.

Research Products

• [Journal Article] Hypoxia Induces an Undifferentiated Phenotype of Oral Keratinocytes in vitro (2015)
  • Author(s): Hiroko Kato, Kenji Izumi, Atsushi Uenoyama, et al.
  • Journal Title: Cells Tissues Organs Volume: 199 Pages: 393-404
  • DOI: 10.1159/000371342
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Hypoxic response of oral mucosa fibroblasts in a 3D collagen lattice (2016)
  • Author(s): Hara Y, Shiomi A, Kato H, Izumi K
  • Organizer: TERMIS-AM
  • Place of Presentation: San Diego, CA, USA
  • Year and Date: 2016-12-11
  • Int'l Joint Research
• [Presentation] A novel approach to identify oral keratinocyte stem cells. (2016)
  • Author(s): Izumi K
  • Organizer: The 39th annual scientific meeting of association for dental sciences of the republic of china
  • Place of Presentation: Taipei, Taiwan
  • Year and Date: 2016-09-02
  • Invited
• [Presentation] Metabolomic profile of human oral keratinocytes under hypoxic culture condition. (2016)
  • Author(s): Kato H, Sugimoto M, Hara Y, Saito N, Shiomi A, Izumi K
  • Organizer: Gordon Research Conference
  • Place of Presentation: Hong Kong, China
  • Year and Date: 2016-08-07
  • Int'l Joint Research
• [Presentation] Metabolomic Profile of Human Oral Keratinocytes Under Hypoxic Culture Condition. (2016)
  • Author(s): Kato H, Sugimoto M, Hara Y, Saito N, Shiomi A, Izumi K:
  • Organizer: International Symposium on Development of Human Resources in Practical Oral Health and Treatment
  • Place of Presentation: Lombok, Indonesia
  • Year and Date: 2016-01-08
  • Int'l Joint Research
• [Presentation] コラーゲンゲル内で3次元培養したヒト口腔粘膜線維芽細胞の低酸素応答 (2016)
  • Author(s): 原 夕子，加藤寛子，塩見 晶，高木律男，泉 健次
  • Organizer: 平成28年度新潟歯学会第2回例会
  • Place of Presentation: 新潟大学歯学部講堂（新潟県 新潟市）
• [Presentation] 異なる培養環境において低酸素が口腔粘膜線維芽細胞の増殖活性に及ばす影響の検討 (2016)
  • Author(s): 原 夕子，加藤寛子，塩見 晶，高木律男，泉 健次
  • Organizer: 第4回低酸素研究会
  • Place of Presentation: 東京女子医科大学先端生命医科学研究所（東京都 新宿区）
• [Presentation] Application of phosphorescent Ir(III) complex for monitoring oxygen levels in a tissue-engineered, ex vivo produced oral mucosa equivalent (EVPOME) - A preliminary study (2015)
  • Author(s): Izumi K, Yoshihara T, Kato H, Hara Y, Saito N, Maeda T, Tobita S
  • Organizer: 第３回低酸素研究会
  • Place of Presentation: 東京女子医科大学(東京都新宿区）
  • Year and Date: 2015-07-25
• [Presentation] Hypoxia induces undifferentiated phenotype of oral keratinocytes in vitro (2014)
  • Author(s): Kato H, Izumi K, Shiomi A, Uenoyama A, Kuo S, Feinberg SE, Maeda T
  • Organizer: TERMIS Americas 2014
  • Place of Presentation: Renaissance Hotel, Washington, DC Downtown, Washington DC, USA
  • Year and Date: 2014-12-13 – 2014-12-16
• [Presentation] 低酸素環境が培養ヒト正常口腔粘膜上皮細胞に及ぼす影響 (2014)
  • Author(s): 泉健次、加藤寛子、塩見晶、上野山敦士、前田健康
  • Organizer: 第12回がんとハイポキシア研究会
  • Place of Presentation: 佐賀県佐賀市ホテルマリターレ創世佐賀
  • Year and Date: 2014-11-21 – 2014-11-22
• [Presentation] 低酸素環境が培養ヒト正常口腔粘膜上皮細胞に及ぼす影響 (2014)
  • Author(s): 泉健次、加藤寛子、塩見晶、上野山敦士、前田健康
  • Organizer: 第2回低酸素研究会
  • Place of Presentation: 東京都新宿区早稲田大学 先端生命医科学センター
  • Year and Date: 2014-07-26